Abstract

DETERMINATION OF COLCHICINE IN HUMAN PLASMA BY A SENSITIVE LC-MS/MS ASSAY

Surya Prakasarao Kovvasu*, Priyanka Kunamaneni, Steven Yeung, Balaji Kodali

ABSTRACT

Colchicine is natural product commonly used to treat gout. The present paper describes the method development and validation of colchicine in human plasma using liquid chromatography–tandem mass spectrometry (LC–MS/MS). Colchicine-d6 was used as an internal standard (IS). Sample extraction was carried out using simple solid phase extraction (SPE). The extracted samples were chromatographed on C18 column using isocratic mobile phase composed of 10 mM ammonium formate (pH 3.5) and methanol 20:80, v/v. The method was linear over the range of 0.04-10.0 ng/mL. A total of five calibration curves were generated during the validation to calculate precision and accuracy. The stability of analyte was extensively evaluated in plasma as well as in aqueous samples and results were met the acceptance criteria defined in US FDA guidelines.

Keywords: Colchicine; Solid–phase extraction (SPE); LC–MS/MS; Method validation.